期刊
JOURNAL OF VIROLOGY
卷 77, 期 2, 页码 841-850出版社
AMER SOC MICROBIOLOGY
DOI: 10.1128/JVI.77.2.841-850.2003
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资金
- NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES [T32AI007325, R01AI025231] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE [P01NS030606, R01NS021954] Funding Source: NIH RePORTER
- NIAID NIH HHS [AI-25231, T32-AI-07325, R01 AI025231, T32 AI007325] Funding Source: Medline
- NINDS NIH HHS [NS-30606, P01 NS030606, NS-21954] Funding Source: Medline
Using isogenic recombinant murine coronaviruses expressing wild-type murine hepatitis virus strain 4 (MRV-4) or MHV-A59 spike glycoproteins or chimeric MHV-4/MRV-A59 spike glycoproteins, we have demonstrated the biological functionality of the N-terminus of the spike, encompassing the receptor binding domain (RBD). We have used two assays, one an in vitro liposome binding assay and the other a tissue culture replication assay. The liposome binding assay shows that interaction of the receptor with spikes on virions at 37degreesC causes a conformational change that makes the virions hydrophobic so that they bind to liposomes (B. D. Zelus, J. H. Schickli, D. M. Blau, S. R. Weiss, and K. V. Holmes, J. Virol. 77: 830-840, 2003). Recombinant viruses with spikes containing the RBD of either MRV-A59 or MHV-4 readily associated with liposomes at 37degreesC in the presence of soluble mCEACAM1(a), except for S4R, which expresses the entire wild-type MHV-4 spike and associated only inefficiently with liposomes following incubation with soluble mCEACAM1(a). In contrast, soluble mCEACAM1(b) allowed viruses with the MHV-A59 RBD to associate with liposomes more efficiently than did viruses with the MHV-4 RBD. In the second assay, which requires virus entry and replication, all recombinant viruses replicated efficiently in BHK cells expressing mCEACAM1(a). In BHK cells expressing mCEACAM1(b), only viruses expressing chimeric spikes with the MHV-A59 RBD could replicate, while replication of viruses expressing chimeric spikes with the MHV-4 RBD was undetectable. Despite having the MHV-4 RBD, S4R replicated in BHK cells expressing mCEACAM1(b); this is most probably due to spread via CEACAM1 receptor-independent cell-to-cell fusion, an activity displayed only by S,R among the recombinant viruses studied here. These data suggest that the RBD domain and the rest of the spike must coevolve to optimize function in viral entry and spread.
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